This technology uses a programmable DNA switch to precisely control enzyme activity, enabling rapid, sensitive detection of diverse targets or selective activation of therapeutic enzymes, making it useful for biosensing and targeted treatments.
The field of biosensing and targeted therapeutics has seen rapid growth due to the increasing need for highly sensitive, specific, and rapid detection of a wide range of biomolecules, as well as the demand for precise activation of therapeutic agents in specific biological contexts. Traditional biosensors and prodrug systems often lack the flexibility and programmability required to detect diverse analytes or to control enzyme activity with high spatial and temporal precision. As diseases become more complex and personalized medicine gains prominence, there is a critical need for platforms that can be easily adapted to new targets, provide amplified and rapid responses, and minimize off-target effects in therapeutic applications.
Current approaches to enzyme regulation and biosensing typically rely on covalent modifications, multi-component assemblies, or complex nanostructures such as DNA origami. These methods can be labor-intensive, difficult to scale, and often require stringent conditions for stability and function. Covalent modification of enzymes may alter their activity or stability, while multi-component systems can suffer from assembly inefficiencies and batch-to-batch variability. Additionally, many existing biosensors are limited to detecting only nucleic acid targets or require thermal cycling, which restricts their use in point-of-care settings. In the context of targeted therapeutics, conventional prodrug strategies may lack the specificity needed to ensure activation only in diseased tissues, leading to potential side effects. These limitations highlight the need for a more versatile, robust, and easily programmable solution for both biosensing and controlled therapeutic activation.
This technology is a modular, programmable DNA-regulated enzyme system that enables precise and reversible control of enzyme activity through an allosteric DNA switch. The core structure consists of an enzyme tethered to its reversible inhibitor via a DNA-based linker that is designed to respond to specific target analytes, such as nucleic acids, proteins, small molecules, or enzymatic activities. In its default state, the inhibitor keeps the enzyme inactive. Upon recognition of the target analyte, the DNA linker undergoes a conformational change—such as hybridization, binding, or cleavage—which disrupts the inhibitor-enzyme interaction and activates the enzyme. This system, known as Single-Molecule DNA Tweezers (SMDTs), is highly versatile: the enzyme, inhibitor, and recognition sequence can be customized for a wide variety of applications, including rapid, isothermal biosensing and targeted therapeutic activation. The technology allows for amplified, sensitive detection of analytes and can function as a prodrug system, selectively activating therapeutic enzymes only in the presence of disease-specific molecular markers.
What differentiates this technology is its unique combination of modularity, programmability, and simplicity. Unlike traditional biosensors or prodrug systems that require complex covalent modifications or multi-component assemblies, this solution operates as a single-molecule, conjugation-free construct. The allosteric DNA switch provides reversible, non-covalent control over enzyme activity, enabling rapid and amplified signal generation without the need for elaborate instrumentation or processing steps. Its programmability allows for easy adaptation to virtually any target analyte by simply modifying the DNA linker sequence or incorporating specific aptamers or substrates. The system’s ability to detect both nucleic acid and non-nucleic acid targets, combined with its isothermal operation and visual or instrument-based readouts, makes it highly suitable for point-of-care diagnostics and precision therapeutics. Additionally, the single-molecule design simplifies synthesis and scalability, offering a practical and broadly applicable platform that stands out from more cumbersome or narrowly focused alternatives.
This technology is available for exclusive licensing. This modular DNA-regulated enzyme system precisely controls enzyme activity via an allosteric DNA switch. An enzyme and its reversible inhibitor are tethered by a DNA sequence, rendering the enzyme inactive. Target molecule binding alters the DNA, disrupting the inhibitor-enzyme interaction and activating the enzyme. This programmable platform functions as a biosensor or prodrug system.
PCT Patent filed: PCT/US2025/034773
Publication: Single-Molecule DNA Tweezers Enable Programmable Control of Enzyme Activity via Arbitrary Molecular Cues